OBJECTIVE:To study the synthesis technology of Ligustilide dipolymer (compound Ⅰ). METHODS:Using phthalic anhydride(compoundⅡ)as raw materials,sodium borohydride was reduced to phthalide(compound Ⅲ),and then syn-thesize ligustilide dipolymer after 4 steps. Ligustilide dipolymer was characterized by 1H-NMR and 13C-NMR. The synthesis technol-ogy of fifth step was optimized by orthogonal test with catalyzer,reaction temperature and reaction time as factors using yield as in-dex. The validation test was conducted. RESULTS:The optimal synthesis technology of fifth step was as follows as aluminium mu-riate of catalyzer,reaction temperature of 150 ℃,reaction time of 5 h. The average yield of validation test was 56.16%(RSD=1.23%,n=3). Total reaction yield was 24.4%,and the purity of target product was 99.4%. CONCLUSIONS:Ligustilide dipoly-mer is synthesized successfully;synthetic process is stable,and its quality is controllable.

BACKGROUND:Fluorescent magnetic nanoparticles with the properties of quantum dots and magnetic particles have good biocompatibility, and can label cels effectively through endocytosis. OBJECTIVE:To validate the feasibility of fluorescent magnetic nanoparticles in labeling human adipose-derived mesenchymal stem cels. METHODS:Healthy human adipose tissue was extracted and adipose-derived mesenchymal stem cels were isolatedin vitro by type I colagenase digestion. Passage 6 cels were incubated with the fluorescent magnetic nanoparticles overnight. Prussian blue staining and laser scanning confocal microscope were used to observe labeled adipose-derived mesenchymal stem celsin vitro after co-culturing with fluorescent magnetic nanoparticles. The tracing effect of labeled adipose-derived mesenchymal stem cels in vivo was detected by fluorescence imaging system. RESULTS AND CONCLUSION:Prussian blue staining showed that the fluorescent magnetic nanoparticles dispersed in the cytoplasm of adipose-derived mesenchymal stem cels in the form of blue particles. Under the laser scanning confocal microscope, the nuclei of adipose-derived mesenchymal stem cels were dyed blue by Hoechest33258, and the cytoplasm was dyed green. The fluorescence imaging results showed that labeled human adipose-derived mesenchymal stem cels had good imaging results. Therefore, as an efficient tracer, the fluorescent magnetic nanoparticles can label human adipose-derived stem cels in vitro and provide a new method for transplantation and transformation of adipose-derived mesenchymal stem cels.

BACKGROUND:Stromal vascular fraction digested from adipose tissue is a mixture of a variety of cel s, and clone-like cel components may have the real ability to maintain the stemness of mesenchymal stem cel s OBJECTIVE:To separate, passage and culture colony-forming cel s in the stromal vascular fraction of adipose tissue, and to test its ability to repair the knee cartilage in animal models. METHODS:Stromal vascular fraction isolated from fresh adipose tissue was cultured to col ect clone-like adherent cel s that were subject to digestion and passage until the third generation. A rabbit model of osteoarthritis was established surgical y, and the third generation of colony-forming cel s were transplantated into the articular cavity of model rabbits. Model rabbits with no cel transplantation served as controls. Cartilage regeneration in the knee joint of model rabbits were observed and compared. RESULTS AND CONCLUSION:At 3 days after transplantation of stromal vascular fraction cel s, clone-like cel colonies appeared in adherent cel s;at 5 days, the amount of clone-like cel colonies and surrounding single cel s were both increased. At the passage 3, the clone-like cel colonies were also visible, which were fusiform in shape. Gross observation and MRI examination showed that these colony-forming cel s could promote cartilage repair in the joint cavity, increase secretion of synovial fluid, and help to control inflammation and narrow ulceration area. Experimental findings are beneficial for optimizing the isolation and culture of adipose-derived mesenchymal stem cel s and provide a new way for the clinical treatment of osteoarthritis.

Aim To investigate the role of substance P in the brain of asthmatic rats.Methods Rats were injected with aluminum hydroxide and OVA allergens to prepare the animal model of asthma. Then the content of c-fos protein in asthmatic rats' brain was detected by immunohistochemical method (SABC).The content of substance P in paraventricular nucleus (PVN), the content of corticotropin-releasing hormone (CRH) in median eminence (ME), and the content of adrenocorticotropic hormone (ACTH) and corticosterone (CORT) in peripheral blood were detected by radioimmunoassay method. Then exogenous SP, SP receptor antagonist S0145 were microinjected in PVN to observe their effect upon lung function and HPA axis in asthmatic rats. Results In asthmatic rats, the content of SP increased within the PVN. The content of CRH, ACTH and CORT decreased (P<0.05).The ratio of expiratory and inspiratory and airway resistance increased. Diaphragm discharge points and lung compliance decreased (P<0.01).After microinject of SP in PVN, there was a further decrease in pulmonary function and the content of CORT, ACTH and CRH in asthmatic rats (P<0.01).While the SP receptor antagonist S0145 might reverse the change of lung function and hypothalamus-pituitary-adrenal axis (HPA axis) in asthmatic rats.Conclusion In asthmatic rats the SP in PVN can affect the function of HPA axis, involved in asthma attacks.

Albendazole and mebendazole were comparatively evaluated in 22 adult patients for their in vivo effects on hookworm eggs. Both drugs were given 200mg twice daily for three consecutive days. Stool specimens wore collected before treatment and during the following five days. The Stool egg count was carried out by Stolls method and each specimen was also cultured by Hara-da-Mori technique for at least eight days. The mean pre-treatment percentage of incubated hookworm eggs that developed to larvae was 75.3% in albendazole group and 68.8% in mebendazole group respectively. One day following the initiation of treatment the mean percentage was remarkably reduced to 0.25% in albeadaole group and no eggs developed to larval stage beyond day 1. 0ne day and two days following the initiation of treatment the mean percentage was 16.23% and 23.13% respectively in mebendazole group. No eggs developed to larval stage on day 3 and thereafter. Albendazole seems to have better ovicidal effect than mebendazole.Albendazole and mebendazole were also comparatively evaluated in 123 adult patients with single or mixed infections of hookworm, ascarisis, and trichuris. A single dose of 400mg was used for both drugs. 2 and 4 weeks after treatment, their stools were examined by brine flotation technique. The hookworm eggs negative conversion ratss 2 weeks after treatment were 78.8% and 26.4% respectively in albeadazole and mebendazole group; while those 4 weeks after treatment were 74.1% and 25.5% respectivley. For ascariasis, the eggs negative conversion rates were 98.0% and 92.0% respectively. For trichuriasis, the negative conversion rates were only 20.6% and 27.7% respectively

BACKGROUND:Large-scale expansion of undifferentiated and multipotential adipose-derived stem cells using serum-free culture system is a difficult issue to be resolved. OBJECTIVE:To establish an in vitro culture system combined with the extracellular matrix in order to investigate the efficiency, effectiveness and security of extracellular matrix on expanding adipose-derived stem cells. METHODS:In vitro isolated adipose-derived stem cells were seeded in traditional two-dimensional plastic plates and extracellular matrix-coated plates supplemented with serum-free medium respectively. After in vitro expansion, total cellnumber, expression of cellsurface markers, cellsenescence degree and multipotent differentiation ability (adipogenic, osteoblastic and chondrogenic differentiation) of adipose-derived stem cells cultured under both conditions were detected and compared. Moreover, the clinical safety of adipose-derived stem cells expanded in extracellular matrix-coated plates was investigated. RESULTS AND CONCLUSION:Total cellnumber of passage 5 adipose-derived stem cells cultured in extracellular matrix-coated plates was 10 times more than that in traditional two-dimensional plastic plates. Flow-cytometric analysis showed that adipose-derived stem cells cultured with extracellular matrix expressed stem cellsurface markers. cellular senescence examination showed that almost al of passage 15 adipose-derived stem cells cultured with extracellular matrix showed no aging, while most passage 5 adipose-derived stem cells cultured by the two-dimensional system aged and lost their proliferation ability. Multidirectional induction of adipose-derived stem cells showed that passage 15 adipose-derived stem cells cultured with extracellular matrix could stil differentiate into adipocytes, osteoblasts and chondrocytes as passage 5 adipose-derived stem cells did, which performed much better than the induced differentiations of passage 5 adipose-derived stem cells cultured by the two-dimensional system. Karyotype analysis and in vivo invasion experiment insured the clinical safety of adipose-derived stem cells expanded with extracellular matrix. Al above results suggest a safe and more efficient expansion system of extracellular matrix for clinical application using the serum-free culture system combined with extracellular matrix.

Objective To study the effect for albendazole therapy for alveolar echinococcus infection in gerbils and mice.Methods Mice and gerbils were infected of metacestode tissues by intraperitoneal (IP) transplantation and treated with albendazole-medicated feeds. The effects were evaluated by comparison of the treated and control groups in terms of host mortality, larval metastases to lungs and liver, final larval weight, histopathological and ultrastructural examination of metacestode tissues. Viability of metacestode tissues at necropsy of treated animals was tested by intraperitoneal transplantation into uninfected animals.Results Albendazole-medicated feeds significantly inhibited larval growth of Echinococcus multilocularis (E. multilocularis) both in mice and gerbils with markedly reduced host mortality and pulmonary and liver metastsaes. Viability test showed that albendazole therapy was parasiticidal in early stage of experimental infection. Light microscopic and ultrastructural examination of metacestode tissues of the treated animals revealed severe destruction and massive necrosis with marked calcification of protoscoleces and residual tissues.Conclusion Continuous long term albendazole therapy in animal models is parasiticidal against larval E. multilocularis especially in early stage of infection.

OBJECTIVETo observe triclabendazole effect on Paragonimus skrjabini in experimentally infected rats,and to develop a new drug for treating paragonimiasis.

METHODSMetacercariae of Paragonimus skrjabini were isolated from crabs (Sinopotamon) collected from endemic area. Wistar rats were infected intraperitoneally. One and two months after infection, they were treated with triclabendazole at the dosage of 300 mg.kg(-1).2 d(-1), 450 mg.kg(-1).3 d(-1) and 600 mg.kg(-1).3 d(-1) respectively. Five patients with Paragonimus skrjabini were treated, with Triclabendazole dosage of 10 mg/kg bid x 3 days.

RESULTSThe worm reduction rates were 50.3%, 80.8% and 86.7% respectively one month after completion of treatment. Dead worms of sesame size recovered from muscles, liver, abdominal cavity, chest cavity and lung were greatly diminished in size and weight in comparison with that of the control group. Many large (about 1 cm) black-colored distended worm cysts were found in the lungs of the control rats. Usually there were two adult worms pairs with numerous eggs in each worm cyst. Most worm cysts in the treated groups of rats were changed into hemorrhagic-necrotic patches. All five patients were cured.

CONCLUSIONTriclabendazole was highly active against Paragonimus skrjabini in rats experimentally infected and patients.

Adult ; Animals ; Anthelmintics ; therapeutic use ; Benzimidazoles ; therapeutic use ; Child ; Female ; Humans ; Male ; Paragonimiasis ; drug therapy ; Parasite Egg Count ; Rats ; Rats, Wistar

Objective To investigate time distribution of pressure of footplate in each stage and vertical anti-force from ground of patients with knee osteoarthritis (KOA).Methods 40 KOA patients, 10 aged people, and 10 common saplings were selected to test time distribution of pressure of footplate in each stage and vertical anti-force from ground in period of monopodia support.Results To unilateral KOA patient, there was not obvious difference to be observed in initial contact phase and forefoot push off phase ( P>0.05), but there was a significant difference in forefoot contact phase and foot flat phase between health side and sick side ( P<0.001), and there was obvious difference decreased buffer time and buffer force coefficient between them ( P<0.01).Conclusion To KOA patients, the time decreases in weight-bearing phase and absorption shocks, and amortization coefficient also decreases. These indexes can be used as the walking index of functional assessment and the objective evidence of curative effect.

Objective To explore the correlation between TCM constitution and sleep status in escort troops. Methods Standardized TCM Constitution Questionnaire and Pittsburgh Sleep Quality Index (PSQI) were used to investigate escort troops. Factors of PSQI and constitution types were analyzed. Results The results of PSQI showed the number of escort troops who had bad sleeping (PSQI>7) was 64, which accounted for 46%of the escort troops, among which, 45 people (70.3%) were justified as biased constitution, while 19 were normal constitution (29.7%). The top three biased constitution in bad sleeping were:qi deficiency, phlegm-dampness and dampness-heat. PSQI factors such as sleep disorder, daytime dysfunction and total score had positive correlation with qi deficiency, yang deficiency, yin deficiency, dampness-heat and blood stasis, and negative correlation with normal constitution. Conclusion Sleep condition of escort troops has correlation with TCM constitution, which indicates that sleep status could be improved by regulating TCM constitution.